Aberrant Epstein–Barr virus persistence in HIV carriers is characterized by anti-Epstein–Barr virus IgA and high cellular viral loads with restricted transcription

Stevens, Servi J.C.; Smits, Paul H.M.; Verkuijlen, Sandra A.W.M.; Rockx, Davy; Gorp, Eric C. M. Van; Mulder, J.; Middeldorp, Jaap M.

doi:10.1097/qad.0b013e3282eeeba0

Cited by 23 publications

(30 citation statements)

References 49 publications

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“…We determined the fractionated non-B-cell populations to be ∼97% pure, suggesting a maximum of ∼3% contaminating B cells. B-cell contamination therefore is unlikely to be the reason we detected BART miRNAs in the non-B-cell fractions, first because infection frequencies are very low, even in immune-suppressed patients with elevated EBV levels (29,30), and second because the viral genome was undetectable in the non-B cells. Overall, our data suggest that in asymptomatic patients BART miRNAs are expressed by latently infected circulating B cells but also are present in noninfected non-B cells, suggesting miRNA transfer in vivo.…”

Section: Ebv Bart Mirnas Are Present In Peripheral B Lymphocytes and mentioning

confidence: 96%

“…Random asymptomatic HIV carriers (n = 197) visiting the Slotervaart Hospital (Amsterdam, the Netherlands) for routine check-up between 2004 and 2006 were enrolled in a previously published study (29). Whole blood (≈10 mL) was collected from these individuals for routine diagnostic testing for plasma HIV-RNA load and CD4 T-cell counts .…”

Section: Methodsmentioning

confidence: 99%

“…The amount, quality, and composition of isolated RNA was analyzed by the NanoDrop 1000 spectrophotometer (Thermo Scientific) for total RNA and an Agilent 2100 Bioanalyzer for small RNA profiles. DNA isolation was performed using a silica-based method described previously (29). The generation of primary (immature) MoDC is explained in detail in SI Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

“…These patients have a restricted EBV-gene-expression pattern similar to that seen in healthy individuals (29). To confirm the cellular tropism of EBV for B cells, we fractionated peripheral blood mononuclear cells (PBMCs) in T-, B-, and non-B-cell populations by cell sorting and measured the EBV-DNA load (DNA copies/mL blood) by quantitative DNA-PCR that targets a conserved region of the viral genome (29).…”

Section: Ebv Bart Mirnas Are Present In Peripheral B Lymphocytes and mentioning

confidence: 99%

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Functional delivery of viral miRNAs via exosomes

Pegtel

Cosmopoulos

Thorley‐Lawson

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

1,424

1,227

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Noncoding regulatory microRNAs (miRNAs) of cellular and viral origin control gene expression by repressing the translation of mRNAs into protein. Interestingly, miRNAs are secreted actively through small vesicles called "exosomes" that protect them from degradation by RNases, suggesting that these miRNAs may function outside the cell in which they were produced. Here we demonstrate that miRNAs secreted by EBV-infected cells are transferred to and act in uninfected recipient cells. Using a quantitative RT-PCR approach, we demonstrate that mature EBV-encoded miRNAs are secreted by EBV-infected B cells through exosomes. These EBV-miRNAs are functional because internalization of exosomes by MoDC results in a dose-dependent, miRNAmediated repression of confirmed EBV target genes, including CXCL11/ ITAC, an immunoregulatory gene down-regulated in primary EBVassociated lymphomas. We demonstrate that throughout coculture of EBV-infected B cells EBV-miRNAs accumulate in noninfected neighboring MoDC and show that this accumulation is mediated by transfer of exosomes. Thus, the exogenous EBV-miRNAs transferred through exosomes are delivered to subcellular sites of gene repression in recipient cells. Finally, we show in peripheral blood mononuclear cells from patients with increased EBV load that, although EBV DNA is restricted to the circulating B-cell population, EBV BART miRNAs are present in both B-cell and non-B-cell fractions, suggestive of miRNA transfer. Taken together our findings are consistent with miRNA-mediated gene silencing as a potential mechanism of intercellular communication between cells of the immune system that may be exploited by the persistent human γ-herpesvirus EBV.

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Section: Ebv Bart Mirnas Are Present In Peripheral B Lymphocytes and mentioning

confidence: 96%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Ebv Bart Mirnas Are Present In Peripheral B Lymphocytes and mentioning

confidence: 99%

See 2 more Smart Citations

Functional delivery of viral miRNAs via exosomes

Pegtel

Cosmopoulos

Thorley‐Lawson

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

1,424

1,227

View full text Add to dashboard Cite

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“…A search for EBV DNA is particularly important in immunocompromised patients with an incomplete humoral response and patients who have received transfusions or immunoglobulins that confound serological test results [28] . It has been reported that immunocompromised patients have higher baseline viral levels than healthy carriers [99,100] , which decline after treatment. A search for EBV DNA is also useful in patients with EBV-related tumors, except for those with AIDS-related brain tumors in whom the blood levels are low because of the bloodbrain barrier [99,101] .…”

Section: Molecular Biologymentioning

confidence: 99%

Serological diagnosis of Epstein-Barr virus infection: Problems and solutions

Paschale

Clerici²

2012

WJV

242

272

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Serological tests for antibodies specific for Epstein-Barr virus (EBV) antigens are frequently used to define infection status and for the differential diagnosis of other pathogens responsible for mononucleosis syndrome. Using only three parameters [viral capsid antigen (VCA) IgG, VCA IgM and EBV nuclear antigen (EBNA)-1 IgG],it is normally possible to distinguish acute from past infection: the presence of VCA IgM and VCA IgG without EBNA-1 IgG indicates acute infection, whereas the presence of VCA IgG and EBNA-1 IgG without VCA IgM is typical of past infection. However, serological findings may sometimes be difficult to interpret as VCA IgG can be present without VCA IgM or EBNA-1 IgG in cases of acute or past infection, or all the three parameters may be detected simultaneously in the case of recent infection or during the course of reactivation. A profile of isolated EBNA-1 IgG may also create some doubts. In order to interpret these patterns correctly, it is necessary to determine IgG avidity, identify anti-EBV IgG and IgM antibodies by immunoblotting, and look for heterophile antibodies, anti-EA (D) antibodies or viral genome using molecular biology methods. These tests make it possible to define the status of the infection and solve any problems that may arise in routine laboratory practice.

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Serological and virological investigation of the role of the herpesviruses EBV, CMV and HHV‐6 in post‐infective fatigue syndrome

Cameron

Flamand

Juwana

et al. 2010

Journal of Medical Virology

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Multiple previous studies have sought evidence for ongoing, active infection with, or reactivation of, Herpesviruses in patients with chronic fatigue syndrome (CFS), with conflicting results. This study aimed to clarify this by studying 20 patients enrolled in a well-characterized model of the onset and evolution of CFS, the prospective cohort of the Dubbo Infection Outcomes Study (DIOS). The patients selected for examination included five CFS patients with primary Epstein-Barr virus (EBV) infection; five CFS patients with acute viral infection not caused by EBV; and 10 matched controls with prompt resolution of primary EBV infection. Serum samples from three timepoints were assayed using a comprehensive range of serological assays for EBV, HHV-6, and CMV. Viral genomes were assessed using quantitative PCR assays. All patients were seropositive for HHV-6, and 10 were seropositive for CMV at infection baseline (five patients and five controls). Low titer CMV IgM antibodies were found at infection baseline in two of these cases and three control patients. HHV-6 IgG antibody titers were highest at infection baseline but did not differ between the CFS cases and the control patients. There were increases in EBV IgG VCA p18, EBNA-1 IgG, and EA IgG titers over time, but these did not differ between CFS cases and control patients. EBV and HHV6 DNA levels were at control levels in a minority of samples, and CMV was undetectable in all samples. These data do not support the hypothesis of ongoing or reactivated EBV, HHV-6, or CMV infection in the pathogenesis of CFS.

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Aberrant Epstein–Barr virus persistence in HIV carriers is characterized by anti-Epstein–Barr virus IgA and high cellular viral loads with restricted transcription

Cited by 23 publications

References 49 publications

Functional delivery of viral miRNAs via exosomes

Functional delivery of viral miRNAs via exosomes

Serological diagnosis of Epstein-Barr virus infection: Problems and solutions

Serological and virological investigation of the role of the herpesviruses EBV, CMV and HHV‐6 in post‐infective fatigue syndrome

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